The Airway Mucus Institute, Yonsei University College of Medicine, Seoul 120-752, Korea.
J Biol Chem. 2011 Sep 30;286(39):34199-214. doi: 10.1074/jbc.M111.247684. Epub 2011 Aug 10.
Mucus hypersecretion is a prominent feature of respiratory diseases, and MUC5B is a major airway mucin. Mucin gene expression can be affected by inflammatory mediators, including prostaglandin (PG) D(2,) an inflammatory mediator synthesized by hematopoietic PGD synthase (H-PGDS). PGD(2) binds to either D-prostanoid receptor (DP1) or chemoattractant receptor homologous molecule expressed on T-helper type 2 cells (CRTH2). We investigated the mechanisms by which PGD(2) induces MUC5B gene expression in airway epithelial cells. Western blot analysis showed that H-PGDS was highly expressed in nasal polyps. Similar results were obtained for PGD(2) expression. In addition, we could clearly detect the expressions of both H-PGDS and DP1 in nasal epithelial cells but not CRTH2. We demonstrated that PGD(2) increased MUC5B gene expression in normal human nasal epithelial cells as well as in NCI-H292 cells in vitro. S5751, a DP1 antagonist, inhibited PGD(2)-induced MUC5B expression, whereas a CRTH2 antagonist (OC0459) did not. These data suggest that PGD(2) induced MUC5B expression via DP1. Pretreatment with extracellular signal-regulated kinase (ERK) inhibitor (PD98059) blocked both PGD(2)-induced ERK mitogen-activated protein kinase (MAPK) activation and MUC5B expression. Proximity ligation assays showed direct interaction between RSK1 and cAMP response element-binding protein (CREB). Stimulation with PGD(2) caused an increase in intracellular cAMP levels, whereas intracellular Ca(2+) did not have such an effect. PGD(2)-induced MUC5B mRNA levels were regulated by CREB via direct interaction with two cAMP-response element sites (-921/-914 and -900/-893). Finally, we demonstrated that PGD(2) can induce MUC5B overproduction via ERK MAPK/RSK1/CREB signaling and that DP1 receptor may have suppressive effects in controlling MUC5B overproduction in the airway.
黏液过度分泌是呼吸道疾病的一个突出特征,MUC5B 是主要的气道黏蛋白。黏蛋白基因的表达可以受到炎症介质的影响,包括前列腺素(PG)D2,一种由造血 PGD 合酶(H-PGDS)合成的炎症介质。PGD2 与 D-前列腺素受体(DP1)或辅助性 T 细胞 2 型趋化因子受体同源物(CRTH2)结合。我们研究了 PGD2 诱导气道上皮细胞中 MUC5B 基因表达的机制。Western blot 分析显示,H-PGDS 在鼻息肉中高度表达。PGD2 的表达也得到了类似的结果。此外,我们可以清楚地检测到鼻上皮细胞中 H-PGDS 和 DP1 的表达,但没有 CRTH2。我们证明 PGD2 可以在体外正常的人鼻上皮细胞和 NCI-H292 细胞中增加 MUC5B 基因的表达。DP1 拮抗剂 S5751 抑制了 PGD2 诱导的 MUC5B 表达,而 CRTH2 拮抗剂(OC0459)则没有。这些数据表明,PGD2 通过 DP1 诱导 MUC5B 的表达。细胞外信号调节激酶(ERK)抑制剂(PD98059)预处理阻断了 PGD2 诱导的 ERK 丝裂原活化蛋白激酶(MAPK)激活和 MUC5B 表达。连接酶检测分析显示 RSK1 和 cAMP 反应元件结合蛋白(CREB)之间的直接相互作用。PGD2 刺激导致细胞内 cAMP 水平增加,而细胞内 Ca2+ 没有这种作用。PGD2 诱导的 MUC5B mRNA 水平通过 CREB 与两个 cAMP 反应元件位点(-921/-914 和-900/-893)的直接相互作用进行调节。最后,我们证明 PGD2 可以通过 ERK MAPK/RSK1/CREB 信号通路诱导 MUC5B 的过度产生,而 DP1 受体可能对控制气道中 MUC5B 的过度产生具有抑制作用。